Speaker
Description
"Neurodevelopmental Disorders (NDDs) include a wide range of conditions characterised by impairments affecting the development of the central nervous system, with implications for cognitive, motor, behavioral, and social functioning.
To tackle the complexity of the mechanisms underlying these dysfunctions suitable experimental models are needed. While animal models and 2D in vitro cultures are limited in recapitulating the complex, human-specific features of neurodevelopment, increasingly sophisticated 3D brain organoids have enabled researchers to study the dynamics of these alterations with unprecedented accuracy, opening new avenues for understanding the regulatory mechanisms of these diseases and testing potential drug therapies.
However, time-consuming procedures and technical challenges still hinder organoids modeling from being scaled-up in a high-reproducible and high-throughput fashion.
We developed a simple and reproducible workflow for the generation and profiling of patient-derived cortical brain organoids (CBOs) at scale. To strengthen the reliability of this pipeline we included over 40 stem cell lines from NDD case-control matched cohorts, along with iPSCs and embryonic stem cell isogenic lines harboring highly penetrant mutations in NDD-related genes.
First, we generated CBOs from iPSC and ESC lines, seeding and culturing them under identical conditions to ensure uniform confluence across all cell lines. Multiple time points were selected to profile the CBOs. At the gene expression level, single-cell data analysis was performed, and immunofluorescence analysis was integrated using an innovative tissue microarray platform and a slide scanner microscope. This approach significantly reduced sectioning, staining, and acquisition time while lowering antibody and reagent costs and minimizing procedural variability. Finally, a dedicated image- analysis pipeline was developed to characterise CBOs by assessing structure and expression using a panel of 16 selected NDD-related biomarkers.
In summary, we present a simplified and cost-effective approach that enhances efficiency, reproducibility, and scalability while minimizing technical variability in CBO characterization. This offers a promising new tool for studying NDD biomarkers.
With applications in disease modeling and drug screening, this study advances our understanding of neurodevelopmental disorders and paves the way for improved therapeutic strategies."
| Author(s) | C. Maderna1*, L. Culotta1*, E Villa1, N. Caporale1,2, D. Ricca1, D. Dalle Nogare1, G. Testa1,2, The Whole Testa Lab |
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| Affiliation(s) | "1 Human Technopole Foundation, Milan, Italy; 2 Department of Experimental Oncology, University of Milan, Milan, Italy" |
